Characterisation of the signal transduction cascade caused by propofol in rat neurons: from the GABA(A) receptor to the cytoskeleton.
The anaesthetic propofol interacts with the GABA(A) receptor, but its cellular signalling pathways are not fully understood. Propofol causes reorganisation of the actin cytoskeleton into ring structures in neurons. Is this reorganisation a specific effect of propofol as apposed to GABA, and which cellular pathways are involved? We used fluorescence-marked actin in cultured rat neurons to evaluate the percentage of actin rings caused by propofol or GABA in combination with rho, rho kinase (ROK), PI3-kinase or tyrosine kinase inhibitors, with or without the presence of extracellular calcium. Confocal microscopy was performed on propofol-stimulated cells and changes in actin between cellular compartments were studied with Western blot. Propofol (3 microg x ml-1), but not GABA (5 microM), caused transcellular actin ring formation, that was dependent on influx of extracellular calcium and blocked by rho, ROK, PI3-kinase or tyrosine kinase inhibitors. Propofol uses rho/ROK to translocate actin from the cytoskeleton to the membrane and its actin ring formation is dependent on an interaction site close to the GABA site on the GABA(A) receptor. GABA does not cause actin rings, implying that this is a specific effect of propofol.
Björnström
K
K
Department of Anaesthesiology, Faculty of Health Sciences, Linköping University, 58185 Linköping, Sweden. karin.bjornstrom-karlsson@imv.liu.se
Turina
D
D
Loverock
A
A
Lundgren
S
S
Wijkman
M
M
Lindroth
M
M
Eintrei
Ch
Ch
D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006993:Hypnotics and Sedatives / Q000494:pharmacology*; D007202:Indicators and Reagents; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009474:Neurons / Q000187:drug effects*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D015742:Propofol / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D011963:Receptors, GABA-A / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects*; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
D000199:Actins; D004791:Enzyme Inhibitors; D006993:Hypnotics and Sedatives; D007202:Indicators and Reagents; D009419:Nerve Tissue Proteins; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D015742:Propofol; D011505:Protein-Tyrosine Kinases; D011963:Receptors, GABA-A; D054460:rho-Associated Kinases